{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T20:20:45Z","timestamp":1790972445108,"version":"4.1.0"},"reference-count":27,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2006,9,25]],"date-time":"2006-09-25T00:00:00Z","timestamp":1159142400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancer Science"],"published-print":{"date-parts":[[2006,11]]},"abstract":"<jats:p>A deoxycytidine analog, gemcitabine (dFdC), is effective for treating solid tumors and hematological malignancies. After being transported into cancer cells, dFdC is phosphorylated to dFdC triphosphate (dFdCTP), which is subsequently incorporated into the DNA strand, thereby inhibiting DNA synthesis. Intracellular dFdCTP is the critical determinant for dFdC cytotoxicity, so therapeutic drug monitoring or <jats:italic>in\u00a0vitro<\/jats:italic> testing of the capability of cancer cells to accumulate dFdCTP may be informative for optimizing dFdC administration. We have developed a new isocratic\u2010elution high\u2010performance liquid chromatography method for quantifying dFdCTP in cancer cells. Samples (500\u00a0\u00b5L) were eluted isocratically using 0.06\u00a0M Na<jats:sub>2<\/jats:sub>HPO<jats:sub>4<\/jats:sub> (pH\u00a06.9) containing 20% acetonitrile, at a constant flow rate of 0.7\u00a0mL\/min and at ambient temperature. Separation was carried out using an anion\u2010exchange column (TSK gel DEAE\u20102SW; 250\u00a0mm\u00a0\u00d7 4.6\u00a0mm inside diameter, particle size 5\u00a0\u00b5L) and monitored at 254\u00a0nm. The standard curve was linear with low within\u2010day and interday variability. The lower detection limit (20\u00a0pmol) was as sensitive as that of the previous gradient\u2010elution method. dFdCTP was well separated from other nucleoside triphosphates. The method could measure dFdCTP in cultured or primary leukemic cells treated <jats:italic>in\u00a0vitro<\/jats:italic> with dFdC. The method was also applicable to simultaneous determination of dFdCTP and cytarabine triphosphate, the results of which demonstrated ara\u2010CTP production augmented by dFdC pretreatment. Thus, our isocratic high\u2010performance liquid chromatography assay method will be of great use because of its sensitivity and simplicity as well as its applicability to biological materials. (<jats:italic>Cancer Sci<\/jats:italic> 2006; 97: 1274\u20131278)<\/jats:p>","DOI":"10.1111\/j.1349-7006.2006.00323.x","type":"journal-article","created":{"date-parts":[[2006,9,25]],"date-time":"2006-09-25T10:22:05Z","timestamp":1159179725000},"page":"1274-1278","source":"Crossref","is-referenced-by-count":12,"title":["A new, simple method for quantifying gemcitabine triphosphate in cancer cells using isocratic high\u2010performance liquid 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